We previously identified a novel, nonselective cation channel in native reactive (type R1) astrocytes (NR1As) from injured rat brain that is regulated by cytoplasmic Ca2+ and ATP (NC(Ca-ATP)) and exhibits sensitivity to block by adenine nucleotides similar to that of sulfonylurea receptor type 1 (SUR1). Here we show that SUR1 is involved in regulation of this channel. NR1As within the site of injury and after isolation exhibited specific binding of FITC-tagged glibenclamide and were immunolabeled with anti-SUR1 antibody, but not with anti-SUR2, anti-Kir6.1 or anti-Kir6.2 antibodies, indicating absence of ATP-sensitive K+ (KATP) channels. RT-PCR confirmed transcription of mRNA for SUR1 but not SUR2. Several properties previously associated exclusively with SUR1-regulated KATP channels were observed in patch-clamp experiments using Cs+ as the charge carrier: (1) the sulfonylureas, glibenclamide and tolbutamide, inhibited NCCa-ATP channels with EC50 values of 48 nm and 16.1 microm, respectively; (2) inhibition by sulfonylureas was lost after exposure of the intracellular face to trypsin or anti-SUR1 antibody; (3) channel inhibition was caused by a change in kinetics of channel closing, with no change in channel amplitude or open-channel dwell times; and (4) the SUR activator ("KATP channel opener"), diazoxide, activated the NCCa-ATP channel, whereas pinacidil and cromakalin did not. Also, glibenclamide prevented cell blebbing after ATP depletion, whereas blebbing was produced by exposure to diazoxide. Our data indicate that SUR1 is functionally coupled to the pore-forming portion of the NC(Ca-ATP) channel, providing the first demonstration of promiscuity of SUR1 outside of the K+ inward rectifier family of channels.

Increasing evidence suggests that miRNAs can act as either tumor suppressors or oncogenes in carcinogenesis. In the present study, we identified the role of miR-34a in regulating telomerase activity, with subsequent effect on cellular senescence and viability. We found the higher expression of miR-34a was significantly correlated with the advanced clinicopathologic parameters in hepatocellular carcinoma. Furthermore, tumor tissues of 75 HCC patients demonstrated an inverse correlation between the miR-34a level and telomere indices (telomere length and telomerase activity). Transient introduction of miR-34a into HCC cell lines inhibited the telomerase activity and telomere length, which induced senescence-like phenotypes and affected cellular viability. We discovered that miR-34a potently targeted c-Myc and FoxM1, both of which were involved in the activation of telomerase reverse transcriptase (hTERT) transcription, essential for the sustaining activity of telomerase to avoid senescence. Taken together, our results demonstrate that miR-34a functions as a potent tumor suppressor through the modulation of telomere pathway in cellular senescence.

ABSTRACT:Fetal drug exposure is determined by the type and concentration of placental transporters, and their regulation is central to the development of new treatments and delivery strategies for pregnant women and their fetuses. We tested the expression of several clinically important transporters in the human placenta associated with various pregnancy conditions (i.e., labor, preeclampsia, and preterm labor-inflammation). Placentas were obtained from five groups of women at the time of primary cesarean section: 1) term no labor; 2) term labor; 3) preterm no labor (delivered for severe preeclampsia); 4) preterm labor without inflammation (PTLNI); and 5) preterm labor with inflammation (PTLI). Samples were analyzed by Western blot and immunohistochemistry to identify changes in protein expression. Relative mRNA expression was determined by quantitative real-time polymerase chain reaction. A functional genomic approach was used to identify placental gene expression and elucidate molecular events that underlie the given condition. Placental expression of ATP-binding cassette transporters from women in labor and women with preeclampsia was unaltered. Multidrug resistance protein 1 (MDR1) and breast cancer resistance protein (BCRP) and mRNA expression increased in placentas of women with preterm labor with inflammation. Molecular pathways of genes up-regulated in PTLI samples included cytokinecytokine receptor interactions and inflammatory response compared with those in the PTLNI group. The mRNA expression of MDR1 and BCRP was correlated with that of interleukin-8, which also increased significantly in PTLI samples. These data suggest that the transfer of drugs across the placenta may be altered in preterm pregnancy conditions associated with inflammation through changes in MDR1 and BCRP.

Objective-Distinct processes govern transition from quiescence to activation during term (TL) and preterm labor (PTL). We sought gene sets responsible for TL and PTL, along with the effector genes necessary for labor independent of gestation and underlying trigger.Methods-Expression was analyzed in term and preterm +/− labor (n =6 subjects/group). Gene sets were generated using logic operations.Results-34 genes were similarly expressed in PTL/TL but absent from nonlabor samples (Effector Set). 49 genes were specific to PTL (Preterm Initiator Set) and 174 to TL (Term Initiator Set). The gene ontogeny processes comprising Term Initiator and Effector Sets were diverse, though inflammation was represented in 4 of the top 10; inflammation dominated the Preterm Initiator Set.Comments-TL and PTL differ dramatically in initiator profiles. Though inflammation is part of the Term Initiator and the Effector Sets, it is an overwhelming part of PTL associated with intraamniotic inflammation.

In the present study, we investigate the effect of curcumin, a major active component isolated from rhizomes of Curcuma longa, on the cytotoxicity of three human carcinoma cell lines (AGS, HT-29 and MGC803) in gastrointestinal tract and a normal gastric epithelial cell line GES-1, and the mechanism of curcumin-induced apoptosis. The results indicated that curcumin inhibited the gastrointestinal carcinoma cell growth in a dose-dependent manner and cytotoxicity was more towards the gastric carcinoma cell AGS and colon carcinoma cell HT-29 compared to normal gastric cell GES-1, and increased externalization of phosphatidylserine residue was observed by Annexin V/PI staining in the two cell lines. Treatment of AGS and HT-29 cells with curcumin enhanced the cleavage of procaspase-3, -7, -8 and -9. Meanwhile, curcumin induced endoplasmic reticulum (ER) stress and mitochondrial dysfunction as evidenced by up-regulation of CCAAT/enhancer binding protein homologous protein (CHOP), phosphorylation of JNK and down-regulation of SERCA2ATPase, release of cytochrome c, decrease of Bcl-2 and reduction of mitochondrial membrane potential in both AGS and HT-29 cells. Overexpression of bax, total JNK, phospho-FADD and total FADD were also observed in curcumin-treated HT-29 cells. Moreover, curcumin decreased cytosolic and ER Ca(2+), but increased mitochondrial Ca(2+) in the two cell lines. 2-Aminoethoxydiphenyl borate, an antagonist of inositol 1, 4, 5-triphosphate receptor, partly blocked curcumin-induced cytosolic Ca(2+) decrease in AGS and HT-29 cells. Additionally, carbonyl cyanide m-chlorophenylhydrazone, an inhibitor of mitochondrial Ca(2+) uptake, reversed curcumin-triggered AGS and HT-29 cells growth inhibition. siRNA to CHOP markedly reduced curcumin-induced apoptosis. These results suggest that curcumin can impact on ER stress and mitochondria functional pathways in AGS and HT-29 cells, death receptor pathway was also involved in curcumin-treated HT-29 cells, thus identifying specific well-defined molecular mechanisms that may be targeted by therapeutic strategies.

Objective: To compare the diagnostic accuracy between cone-beam computed tomography (CBCT) and periapical radiography for detecting simulated external apical root resorption (EARR) in vitro. Materials and Methods: The study sample consisted of 160 single-rooted premolar teeth for simulating EARR of varying degrees according to four setups: no (intact teeth), mild (cavity of 1.0 mm in diameter and depth on root surface), moderate (0.4 mm, 0.8 mm, 1.2 mm, and 1.6 mm root shortening), and severe (2.4 mm, 2.8 mm, 3.2 mm, and 3.6 mm root shortening). Two groups of radiographic images were obtained via CBCT and periapical radiography. The absence or presence and the severity for all resorption lesions were evaluated blindly by two calibrated observers. Results: With the CBCT method, the rates of correct classification of no, mild, moderate, and severe EARR were 96.3%, 98.8%, 41.3%, and 87.5%, respectively; with the periapical radiography method, the rates were 82.5%, 41.3%, 68.8%, and 92.5%, respectively. Highly significant differences were found between the two imaging methods for detection of mild (P , .001), moderate (P , .001), and all EARR (P , .001). For detection of all EARR, the sensitivity and specificity values were 75.8% and 96.3% for CBCT, compared with 67.5% and 82.5% for periapical radiography. Conclusion: CBCT is a reliable diagnostic tool to detect simulated EARR, whereas periapical radiography underestimates it. However, if a periapical radiograph is already available to the diagnosis of EARR, CBCT should be used with extreme caution to avoid additional radiation exposure. (Angle Orthod. 2013;83:189-195.)

Local annual multi-professional training as provided by PROMPT was temporally associated with improved obstetric outcomes.

Intrauterine hypoxia impacts fetal growth and organ function. Inducible nitric oxide synthase (iNOS) and neuronal NOS (nNOS) expression was measured to assess the response of fetal hearts to hypoxic (HPX) stress. Pregnant guinea pigs were housed in a hypoxic chamber (10.5% O 2 for 14 d, n ϭ 17) or room air [normoxic (NMX), n ϭ 17͔. Hearts of anesthetized near-term fetuses were removed. mRNA ͓hypoxia-inducible factor, (HIF)-1␣, 1␤, 2␣, 3␣, iNOS, and nNOS͔ and protein levels (HIF-1␣, iNOS, and nNOS) of fetal cardiac left ventricles were quantified by real time polymerase chain reaction (PCR) and Western analysis, respectively. Cardiac nitrite/nitrate levels were measured in the presence/absence of L-N 6 -(1-iminoethyl)-lysine (L-NIL), an iNOS inhibitor, administered to pregnant sows. Hypoxia significantly increased fetal cardiac HIF-1␣ and -2␣ mRNA, HIF-1␣ protein but not HIF-3␣ or -1␤ mRNA levels. Hypoxia increased both iNOS mRNA (by 5ϫ) and protein (by 23%) levels but had no effect on nNOS levels. Nitrite/nitrate levels were increased in HPX hearts by 2.5ϫ and decreased with L-NIL by 67 Ϯ 14%. Thus, up-regulation of iNOS-derived nitric oxide (NO) generation is an important mechanism by which fetal hearts respond to chronic hypoxic stress. T he adaptive response of the fetal heart to intrauterine stress is critical for its survival. Several studies using high altitude (1,2) exposure to acute and chronic hypoxia (2,3), and anemia (4) have demonstrated how the fetal cardiovascular system responds to hypoxic (HPX) stress. Depending on the severity and duration of the HPX conditions, as well as the gestational age of the fetus, cardiac adaptations have been associated with altered coronary blood flow (2,5,6), increased heart size (1), and decreased contractile performance (7). The underlying mechanisms mediating these changes in fetal heart morphology and function are not fully understood.Hypoxia is a potent stimulus for gene activation of several genes (8), including nitric oxide synthase (NOS), the synthetic enzyme that generates nitric oxide (NO) from L-arginine oxidation (9). NO is derived from three isoforms of NOS ͓endothelial NOS (eNOS); neuronal NOS (nNOS); and inducible NOS (iNOS)͔, all of which are expressed in the heart (10,11). Specifically, eNOS is expressed constitutively in both endothelial cells and cardiomyocytes (10), nNOS in both cardiomyocytes and conducting tissue of cardiac ventricles (12), and iNOS in cardiomyocytes (12) and in mature hearts under conditions of hypoxia (12-14), heart failure (15), left ventricular hypertrophy (16) and cardiac cyanosis in children (17). NOS gene expression has been reported to be oxygensensitive, with levels varying among cardiac cells (18,19) and endothelial cells of differing vascular origin (20 -22). For example, hypoxia increases eNOS expression in porcine coronary artery endothelial cells (20), newborn (23-25) and adult rabbit heart ventricles (26), and adult guinea pig ventricles (27), and decreases expression in pulmonary artery endothelial cell...